A case report of serological evidence of paramyxoviruses related to Porcine orthorubulavirus in Mexican bats

In this report, we showed the presence of antibodies to Porcine orthorubulavirus (PRV) in Mexican bats using a serological approach. A total of 42 bats, belonging to seven different species, were sampled from two different refuges/caves, located near to a pig fattening area where spontaneous outbreaks of PRV had occurred. Analysis by serum-virus neutralizing and immunoperoxidase monolayer assay revealed the presence of antibodies in fifteen out of 42 investigated bats (i.e. 35%), six of them were also positive by Paramyxoviridae family using PCR assay targeting the L gene of paramyxoviruses. This case demonstrates for the first time antibodies detection of this virus in different bats species which is important for our understanding of PRV ecology, evolution and mechanism of cross-species transmission. These findings support the hypothesis that bats could act as an intermediate or natural host for interspecies transmission of certain paramyxoviruses

Immunogenicity of a recombinant hemagglutinin neuraminidase-Porcine rubulavirus produced by Escherichia coli of Porcine rubulavirus gives protective immunity of litter after challenge

Porcine rubulavirus (PRV) is a contagious virus that affects the Mexican swine industry. This work aimed to evaluate the immunogenicity of an recombinant hemagglutinin neuraminidase-Porcine rubulavirus (rHN-PorPV) candidate vaccine on pregnant sows, and the protective efficacy afforded to their 7-day- old suckling piglets against PRV lethal challenge. Three sows were immunized with rHN-PorPV formulated with immune-stimulating complex (ISCOMs) and two sows with rHN- PorPV protein alone as well as a mock-immunized pregnant sow (negative control). Quantitative ELISA detected a high concentration of anti-rHN-PorPV Immunoglobulin G (IgG) antibodies in sow sera after the second dose of vaccine administered on day 14 until farrowing, showing viral-neutralizing and cross-neutralization activity against different variants of PRV. Sera samples from piglets of immunized sows (with or without adjuvant), showed high concentrations of IgG antibodies. As expected, piglets from the negative control sow (n=5), exhibited severe signs of disease and 100% of mortality after PRV challenge study. Conversely, 75% and 87.5% of the piglets born from the rHN-PorPV and the rHN-PorPV-ISCOMs-immunized sows (n= 8), survived, respectively, showing milder PRV clinical signs. Our data indicate that rHN-PorPV candidate vaccine produced in Escherichia coli induces efficient humoral response in pregnant sows and that the maternally derived immunity provides high protection to suckling piglets against PRV lethal challenge

Familial hypercholesterolaemia in children and adolescents from 48 countries: a cross-sectional study

Background: Approximately 450 000 children are born with familial hypercholesterolaemia worldwide every year, yet only 2·1% of adults with familial hypercholesterolaemia were diagnosed before age 18 years via current diagnostic approaches, which are derived from observations in adults. We aimed to characterise children and adolescents with heterozygous familial hypercholesterolaemia (HeFH) and understand current approaches to the identification and management of familial hypercholesterolaemia to inform future public health strategies. Methods: For this cross-sectional study, we assessed children and adolescents younger than 18 years with a clinical or genetic diagnosis of HeFH at the time of entry into the Familial Hypercholesterolaemia Studies Collaboration (FHSC) registry between Oct 1, 2015, and Jan 31, 2021. Data in the registry were collected from 55 regional or national registries in 48 countries. Diagnoses relying on self-reported history of familial hypercholesterolaemia and suspected secondary hypercholesterolaemia were excluded from the registry; people with untreated LDL cholesterol (LDL-C) of at least 13·0 mmol/L were excluded from this study. Data were assessed overall and by WHO region, World Bank country income status, age, diagnostic criteria, and index-case status. The main outcome of this study was to assess current identification and management of children and adolescents with familial hypercholesterolaemia. Findings: Of 63 093 individuals in the FHSC registry, 11 848 (18·8%) were children or adolescents younger than 18 years with HeFH and were included in this study; 5756 (50·2%) of 11 476 included individuals were female and 5720 (49·8%) were male. Sex data were missing for 372 (3·1%) of 11 848 individuals. Median age at registry entry was 9·6 years (IQR 5·8-13·2). 10 099 (89·9%) of 11 235 included individuals had a final genetically confirmed diagnosis of familial hypercholesterolaemia and 1136 (10·1%) had a clinical diagnosis. Genetically confirmed diagnosis data or clinical diagnosis data were missing for 613 (5·2%) of 11 848 individuals. Genetic diagnosis was more common in children and adolescents from high-income countries (9427 [92·4%] of 10 202) than in children and adolescents from non-high-income countries (199 [48·0%] of 415). 3414 (31·6%) of 10 804 children or adolescents were index cases. Familial-hypercholesterolaemia-related physical signs, cardiovascular risk factors, and cardiovascular disease were uncommon, but were more common in non-high-income countries. 7557 (72·4%) of 10 428 included children or adolescents were not taking lipid-lowering medication (LLM) and had a median LDL-C of 5·00 mmol/L (IQR 4·05-6·08). Compared with genetic diagnosis, the use of unadapted clinical criteria intended for use in adults and reliant on more extreme phenotypes could result in 50-75% of children and adolescents with familial hypercholesterolaemia not being identified. Interpretation: Clinical characteristics observed in adults with familial hypercholesterolaemia are uncommon in children and adolescents with familial hypercholesterolaemia, hence detection in this age group relies on measurement of LDL-C and genetic confirmation. Where genetic testing is unavailable, increased availability and use of LDL-C measurements in the first few years of life could help reduce the current gap between prevalence and detection, enabling increased use of combination LLM to reach recommended LDL-C targets early in life

Studies of the molecular genetics and epidemiology of porcine rubulavirus infection

Porcine rubulavirus (PorPV) infection emerged spontaneously in pigs in Mexico in the early 1980s. Since the report of the initial outbreak of the disease, only one full-length genome from a strain isolated in 1984 (PorPV-LPMV/1984) has been sequenced. There is therefore limited information about the genetic variation of this virus. The overall objective of this thesis was to develop molecular techniques to help in the diagnostic field and to investigate in greater detail the full genomes of several isolates, and if possible, gain insights into the persistence, molecular epidemiology and the possible reservoirs of PorPV. In addition, a characterisation of the immune response during acute and persistent infection was included. A real-time RT-PCR was developed for the detection of viral RNA from PorPV in clinical samples using TaqMan technology and primers for the P gene. This assay was highly sensitive (approximately 10 copies per reaction), specific, reproducible and a very useful tool for molecular diagnostics and for enabling studies of various aspects of PorPV throughout this thesis. RT-PCRs based on the NP and P genes were used to study the tissue distribution of the virus. Viral mRNA in the lymph nodes showed that the NP gene was consistently detected in the parotid, submaxilar, cervical and mesenteric nodes and the pancreas. Full-length genomes were sequenced from new isolates obtained from clinical cases of infected swine. The genetic comparison and phylogenetic analysis indicated that three different genetic variants of PorPV had spread in the swine population and that a new generation of circulating virus with a pronounced attenuation has begun to emerge in nature. We also report the isolation of PorPV, or a related virus, from frugivorous, insectivorous, and hematophagous bats. A partial genome sequence analysis showed a 99.97 - 100% amino acid identity to the reference strain isolated from swine. However, larger parts of the genome must be sequenced to ascertain the genetic relationship between these viruses. The study of the immune response during acute and persistent infection revealed enhanced levels of CD8+, CD4+ and CD2+ T-cells in all infected pigs at 10 days PI. CD8+ T-cell subpopulations were significantly higher (p<0.05) at 10 and 250 days PI, and CD4+ T-lymphocytes were also significant at 250 days PI. In summary, this work developed molecular techniques that can be used to study the pathogenesis and molecular epidemiology of PorPV. The knowledge of the presence of different virus variants in nature, associated with a wildlife reservoir of PorPV can provide greater knowledge regarding the molecular genetic changes and useful data to establish new strategies in the control of this virus in Mexico

Development of Novel Recombinant Antigens of Nucleoprotein and Matrix Proteins of Porcine orthorubulavirus: Antigenicity and Structural Prediction

Blue eye disease (BED) is a swine viral infection that affects the pork industry of Mexico. Porcine orthorubulavirus (PRV) is the etiological agent, and the hemagglutinin-neuraminidase protein (HN) is characterized as the best antigen for serological tests, although other structural proteins, including the nucleoprotein (NP) and the matrix (M) protein, have been investigated during the infection of members of the Paramyxoviridae family, generating promising results. Herein, for the first time, we successfully produced and characterized both the NP and M proteins of PRV by using a recombinant strategy in the E. coli heterologous system. The ORF of the NP and M genes were cloned in-frame with the pET-SUMO expression vector. Recombinant proteins proved to be a sensitive target to detect seroconversion at 7 days until 28 days in vaccinated mice (BALB/c) by indirect ELISAs. Immunoreactivity was also tested using porcine serum samples, in which antibodies were recognized from early stages to a persistence of PRV infection, which is indicative that these proteins contain properties similar to native antigens. The predicted tertiary structure showed that both proteins have a conserved structure that resembles those found in others Paramyxovirus. Our results pave the way for developing biotechnological tools based on these proteins for the control and prevention of BED

Tópicos selectos de ciencias químicas

La Química es una de las ramas de la ciencia que ha tomado mayor importancia en las últimas décadas. Es imposible no pensar en ella al escuchar los avances milagrosos de fármacos, materiales inteligentes, tecnología nanométrica, computadoras cuánticas o procesos catalíticos, donde la transformación controlada de la materia ha logrado la creación de sustancias con propiedades hechas a medida. En ese sentido, el papel de la Facultad de Química, en la formación de profesionales de dicha área, cobra una importancia superlativa; además del compromiso y la responsabilidad de formar sujetos no sólo con conocimiento, sino desarrollar su conciencia social y ecológica, indispensables para la situación actual y futura.Universidad Autónoma del Estado de México